CN111936167A

CN111936167A - Improvement of CD47 blocking therapy by EGFR antibodies

Info

Publication number: CN111936167A
Application number: CN201980021372.3A
Authority: CN
Inventors: G·H·Y·林; N·N·维尔勒; M·M·王; R·A·乌格
Original assignee: Trillium Therapeutics ULC
Current assignee: Pfizer Inc
Priority date: 2018-03-13
Filing date: 2019-03-08
Publication date: 2020-11-13
Also published as: EP3765080A1; AU2019235627A1; CA3093549A1; JP2021515779A; US20210040219A1; EP3765080A4; WO2019173903A1

Abstract

CD47+ disease cells, such as cancer cells, are treated using a CD47 blocker in combination with an EGFR antibody, such as cetuximab. The anti-cancer effect of cetuximab is enhanced in the presence of sirpa Fc. Particular combinations include sirpa Fc forms that comprise Fc that is an IgG1 or preferably an IgG4 isotype. These combinations are particularly useful in the treatment of solid tumors and hematological cancers, including lymphomas, leukemias, and myelomas.

Description

Improvement of CD47 blocking therapy by EGFR antibodies

Technical Field

The present disclosure relates to methods and uses of drugs that block the CD 47/SIRPa interaction. More particularly, the disclosure relates to methods and uses that, in combination, can be used to improve cancer therapy.

Background

Cancer cells are targeted for destruction by antibodies that bind to cancer cell antigens and through Fc receptor binding to the Fc portion of the antibodies to recruit and activate macrophages. The binding between CD47 on cancer cells and sirpa on macrophages signals "do't eat me" that many tumor cells are able to escape destruction by macrophages. It has been shown that inhibition of the CD 47/sirpa interaction (CD47 blockade) will "see" the macrophages and destroy the target CD47+ cancer cells. The use of sirpa to treat cancer by CD47 blockade is described in WO 2010/130053.

WO2014/094122 to Trillium Therapeutics describes a protein drug that inhibits or antagonizes the interaction between CD47 and sirpa. This CD47 blocker is a form of human sirpa that incorporates specific regions of its extracellular domain linked to a particularly useful form based on the Fc region of IgG 1. In this form, sirpa Fc drugs show a significant effect on the viability of cancer cells presenting with the CD47+ phenotype. This effect is seen in particular on Acute Myeloid Leukemia (AML) cells and many other types of cancer. Soluble forms of SIRP with significantly altered primary structure and potent CD47 binding affinity are described in WO 2013/109752.

Other CD47 blockers have been described, and these include various CD47 antibodies (see, e.g., US8562997 to Stanford and WO2014/123580 to InhibRx), each of which contains a different antigen binding site, but collectively have the ability to compete with endogenous sirpa for binding to CD47, interact with macrophages, and ultimately increase CD47+ disease cell depletion. These CD47 antibodies have in vivo activities that are quite different from the activities inherent to drugs that incorporate sirpa structures. For example, the latter showed negligible binding to erythrocytes, while the opposite properties of the CD47 antibody and the high affinity sirpa variant require a strategy to accommodate the drug "pool" after administration.

Other agents have also been proposed to block the CD47/SIRP α axis. These agents include the CD47Fc protein described in WO2010/083253 of Viral Logic, as well as SIRPa antibodies as described in WO2013/056352 of University Health Network, US 6913894 of Eberhard, and elsewhere.

The CD47 blocking method has great clinical prospect in the development of anti-cancer drugs. There is a need to provide methods and means to improve the effects of these drugs, and in particular to exploit the effects of CD47 blockers that incorporate the CD 47-binding form of sirpa.

Disclosure of Invention

The anti-cancer effect of anti-tumor antibodies is improved when combined with a CD47 blocker. More specifically, the anti-cancer effect of Epidermal Growth Factor Receptor (EGFR) antibodies is improved when combined with a CD47 blocker in the form of sirpa Fc. The present disclosure demonstrates that the anti-cancer effect of EGFR antibodies is enhanced when administered in combination with sirpa Fc. In embodiments, the sirpa Fc has an IgG4 isotype and comprises an IgV domain of human sirpa, and the EGFR antibody is cetuximab (cetuximab). The enhancement of cetuximab activity by sirpafc is manifested, for example, by an increased depletion of treated EGFR + cancer cells, a decreased tumor growth rate, and/or an enhanced survival of the treated subject, as compared to the results of either agent alone.

In one aspect, there is provided a method for treating a subject presenting with CD47+ disease cells, comprising administering to the subject a combination comprising an IgG4 isotype of sirpafc (referred to as sirpafc 4) and an EGFR antibody, such as cetuximab, including commercially available forms thereof,

in a related aspect, there is provided the use of sirpa G4 in combination with an EGFR antibody for treating a subject having CD47+ disease cells (such as cancer).

In another aspect, a pharmaceutical combination comprising sirpa G4 and an EGFR antibody is provided for use in treating CD47+ disease cells.

In another aspect, a kit is also provided comprising a pharmaceutical combination comprising sirpa G4 and an EGFR antibody, together with instructions teaching their use for treating CD47+ disease cells.

In a specific embodiment, the combination of a CD47 blocker and an EGFR antibody is used to treat a solid tumor or a hematological cancer, such as myeloma, lymphoma or leukemia.

In an alternative embodiment, the sirpafc used in combination with an EGFR antibody is sirpag 1. In other alternative embodiments, the EGFR antibody is panitumumab (panitumumab) or a commercially available form thereof

In a related aspect, the combination of the invention is used to treat EGFR + cancer cells, including EGFR + and CD47+ cancer cells.

Other features and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from this detailed description.

Drawings

Figure 1 shows that increased tumor growth inhibition (a and C) and improved survival (B) were observed when Fadu (human head and neck tumor cell line) tumor-bearing mice were treated with the anti-EGFR antibody cetuximab (3mg/kg, 2 times per week for 2 weeks) in combination with sirpa G4(10mg/kg, 5 times per week for 6 weeks) starting on day 3 post tumor vaccination compared to either the anti-EGFR antibody cetuximab (3mg/kg, 2 times per week for 2 weeks) monotherapy or sirpa G4(10mg/kg, 5 times per week) monotherapy.

Detailed Description

The present disclosure provides methods, uses, combinations and kits useful for treating a subject presenting with disease cells having the CD47+ phenotype. In embodiments, the disease cells have the phenotype of CD47+ and EGFR +. In this method, a CD47+ cancer subject receives a combination of an EGFR antibody (such as cetuximab) and a CD47 blocker, the latter preferably being an Fc fusion form of human sirpa, i.e., sirpa Fc, wherein the Fc is preferably an IgG4 isotype or an Fc receptor binding variant thereof, referred to as sirpa G4. The effect of EGFR antibodies was significantly enhanced by CD47 binding to sirpa G4. Therapeutic effects are evident when the CD47+ disease cells are CD47+ cancer cells and tumors that have a phenotype that is also CD47+, and preferably EGFR + cancer cells.

The term "CD 47 +" is used to indicate the phenotype of a cell targeted by the binding of the CD47 blockers of the present invention. Cells of CD47+ can be identified by flow cytometry using the CD47 antibody as an affinity ligand. Appropriately labeled CD47 antibodies are commercially available for this use (e.g., the antibody product of clone B6H12 is available from BD Biosciences). Cells phenotypically examined for CD47 may include standard tumor biopsy samples, including in particular blood samples taken from subjects suspected of carrying endogenous CD47+ cancer cells. CD47 disease cells of particular interest as targets for therapy using the drug combinations of the present invention are those that "overexpress" CD 47. These CD47+ cells are typically disease cells and present on their surface a CD47 density that exceeds the normal CD47 density of a given type of cell. CD47 overexpression will vary between different cell types, but refers herein to any level of CD47, as determined, for example, by flow cytometry or by immunostaining or by gene expression analysis, etc., that is greater than the level measurable on a corresponding cell having a CD47 phenotype that is normal for that cell type.

The term "CD 47+ disease cells" thus refers to cells associated with disease and having the phenotype CD47 +. In one embodiment, the CD47+ disease cell is a cancer cell.

In embodiments, the CD47 blocker is an IgG4 version of human sirpa Fc, which CD47 blocker interferes with and inhibits or blocks signal transmission caused when CD47 interacts with sirpa. As described in WO2014/094122 to Trillium Therapeutics, the entire contents of which are incorporated herein by reference, a preferred sirpa G4 is an Fc fusion form of a region of human sirpa that interacts with CD47 and has been shown to have anti-cancer activity. As used herein, the term "human sirpa" refers to the wild-type endogenous mature form of human sirpa. In humans, sirpa proteins exist in two major forms. One form, variant 1 or V1 form, has the amino acid sequence designated NCBI RefSeq NP _542970.1 (residues 27-504 constitute the mature form). The other form, variant 2 or V2 form, differs by 13 amino acids and has the amino acid sequence denoted CAA71403.1 in GenBank (residues 30-504 constitute the mature form). Both forms of sirpa account for about 80% of the sirpa forms present in humans, and both are encompassed herein by the term "human sirpa". The present disclosure most particularly relates to pharmaceutical combinations comprising human SIRP variant 2 form or V2.

In the pharmaceutical combination of the invention, a sirpa Fc fusion protein has a sirpa component (106-mer) that includes at least residues 32-137 of human sirpa, which constitutes and defines an IgV domain of the form V2 according to the current nomenclature. As shown below, this SIRP α sequence is referred to herein as SEQ ID No. 1.

In a preferred embodiment, the sirpa Fc fusion protein incorporates the IgV domain as defined in SEQ ID No.1, and additional flanking residues that are contiguous within the sirpa sequence. This preferred form of IgV domain represented by residues 31-148 of the V2 form of human sirpa is a 118-mer having the sequence shown below:

sirpa Fc proteins incorporate an Fc region with effector functions. Fc refers to a "crystallizable fragment" and refers to the constant region of an antibody, which primarily includes components within the heavy chain constant region and hinge region. In embodiments, the Fc region includes a lower hinge-CH 2-CH3 domain. More preferably, the Fc region comprises the CH1-CH2-CH3 domains.

An Fc component with "effector function" is an Fc component with at least some native or engineered function, such as at least some contribution to antibody-dependent cellular cytotoxicity or some ability to fix complement. Likewise, Fc will bind at least to Fc receptors.

In embodiments, the Fc region comprises the sequence of a wild-type human IgG4 constant region. In an alternative embodiment, the Fc region incorporated into the fusion protein is derived from any IgG4 antibody having a constant region with effector activity that is present but is naturally significantly less potent than the IgG1 Fc region. The sequence of such Fc region may, for example, correspond to the Fc region of any of the following IgG4 sequences: p01861 (residues 99-327) from UniProtKB/Swiss-Prot and CAC20457.1 (residues 99-327) from GenBank. In a particular and preferred embodiment, the G4 Fc region incorporates an alteration at position 228(EU numbering) in which the serine at this position is substituted with proline (S228P), thereby to stabilize the disulfide bond within the Fc dimer.

In a specific embodiment, the Fc region is based on the amino acid sequence of human IgG4, which is denoted as P01861 in UniProtKB/Swiss-Prot, residues 99-327, and has the amino acid sequence shown below, and said sequence is referred to herein as SEQ ID No. 6:

in an alternative embodiment, SIRP α Fc has an Fc region based on the amino acid sequence of human IgG1, which is denoted as P01857 in UniProtKB/Swiss-Prot, residues 104 and 330, and has the amino acid sequence shown below:

in a particular embodiment, when the Fc component is IgG4 Fc, the Fc incorporates at least S²²⁸P mutation and has the amino acid sequence as listed below, and the sequence is referred to herein as:

in a specific and preferred embodiment, the sirpa Fc fusion protein has an amino acid sequence as shown below: in this embodiment, the Fc component of the fusion protein is based on IgG4 and incorporates S²²⁸And (4) P mutation.

This sirpafc fusion protein is referred to as sirpag 4.

In an alternative embodiment, a sirpa Fc fusion protein has an amino acid sequence as shown below: in this embodiment, the Fc component of the fusion protein is based on IgG 1:

this sirpafc fusion protein is referred to as sirpag 1.

In a preferred embodiment, a sirpafc protein is provided and used in a secreted homodimeric fusion form, wherein the two copies of the fusion protein are coupled by covalent binding between cysteines present in separate sirpafc single polypeptide chains (e.g., sirpafc 4 chain having SEQ ID No. 6).

The pharmaceutical combination of the invention comprises SIRP α G4 or SIRP α G1 as just described, and an agent, such as an antibody, which binds to the human epidermal growth factor receptor (hEGFR), a protein that is present on the surface of many different cell types, including in particular skin cells, such as keratinocytes. As used herein, the term "hEGFR" (EGFR +) refers to any protein that includes the product of the human her-1 gene expressed and processed, wherein the protein is referred to as UniProtKB/Swiss-Prot P00533. The term EGFR is used generically herein and refers to the wild-type protein and all naturally occurring variants thereof. The term "wtEGFR" is more specifically used only to indicate the wild-type form of human EGFR. The term "EGFRvIII" refers to an EGFR variant protein that comprises the product of expression and processing of the her-1 gene variant lacking exons 2-7, and thus comprises only the polypeptide sequences encoded by exons 1 and 8 of her-1. EGFR + disease cells are those that will bind cetuximab or any other antibody selective for EGFR binding.

For the purpose of identifying disease cells that can be targeted by the EGFR antibodies of the present invention, the commercial test EGFR pharmdx (dako) is conveniently used. This is a semi-quantitative immunohistochemical assay for determining EGFR protein overexpression in colorectal tissues. Positive or negative results help to classify abnormal cells/tissues and provide the basis for selection of EGFR + tumors.

The combinations of the present invention are more particularly and in one embodiment based on the trade name now available

hEGFR antibody, known as cetuximab, is commercially available from Eli Lilly and Company. Cetuximab is a recombinant human/mouse chimeric IgG1 antibody that specifically binds to the extracellular domain of wtEGFR. Amino acid sequences of the complete heavy chain (SEQ ID No.8) and the complete light chain (SEQ ID No.9) of cetuximab.

In one embodiment, sirpa G4 is combined with formulated cetuximab or commercially formulated

Are used in combination.

Sold as a sterile clear colorless liquid at pH 7.0 to 7.4, which may contain small amounts of readily visible white amorphous particles of cetuximab. Each disposable 50mL vial contained 100mg cetuximab at a concentration of 2mg/mL and was formulated as a preservative-free solution containing 8.48mg/mL sodium chloride, 1.88mg/mL disodium hydrogen phosphate heptahydrate, 0.42mg/mL sodium dihydrogen phosphate monohydrate, and water for injection USP.

In an alternative embodiment, SIRP α G4 is compared to that currently commercially available and sold under the trade name SIRP α G4

EGFR antibodies known as panitumumab are sold for use in combination. Panitumumab is a recombinant full-length human IgG2 antibody that specifically binds to the extracellular domain of wtEGFR. The amino acid sequences of the heavy and light chains of panitumumab are listed in US6,235,883 and US 7,807,798, which are incorporated herein by reference.

Sold as a sterile, colorless, preservative-free solution containing 20mg/mL Vectibix (panitumumab) in a disposable vial, e.g., a 5mL disposable vial containing 100mg panitumumab (20mg/mL) in 5 mL; a10 mL disposable vial contained 200mg panitumumab (20mg/mL) in 10 mL; and each 20mL disposable vial contained 400mg panitumumab in 20mL (20mg/mL)

Each of the drugs contained in the pharmaceutical combination of the present invention may be formulated separately for combined use. These drugs are said to be used "in combination" when the effects of each drug overlap in the recipient of the two drugs, and when the two drugs are provided in combination in an injectable physical mixture or in a separately packaged form, such as in a kit. The combination may be generated by administering an EGFR antibody to a subject, followed by SIRPaFc, and vice versa.

The two drugs in the combination act synergistically such that sirpa G4 enhances the effect of cetuximab on target cells, and in particular on target cancer cells of CD47+ and EGFR +. This benefit is manifested as a statistically significant improvement in a given target cell or tumor suitability or viability parameter. For example, the benefit in CD47+/EGFR + cancer cells when exposed to a combination of a CD47 blocker and an EGFR antibody can be a statistically significant reduction in the number of viable cancer cells (and thus depletion) relative to untreated or single agent treatment, or a reduction in the number of cancer cells or size/volume of the tumor, or an improvement in the endogenous location or distribution of any particular tumor type, or an enhancement in survival parameters. In embodiments, the improvement resulting from the drug combination treatment appears to be at least additive and synergistic as desired relative to results obtained using sirpa G4 alone or cetuximab alone.

Specifically, sirpa Fc, such as sirpa G4, for example, may increase the efficacy of cetuximab against cetuximab-resistant cancers or cells that develop resistance to cetuximab therapy. Sirpa G4 may also increase the efficacy of cetuximab on cells with lower levels of EGFR by recruiting ADCC/ADCP mechanisms. Sirpa G4 may also increase the efficacy of cetuximab on cells that develop resistance to cetuximab by up-regulating alternative signaling pathways, such as increasing ERBB2 expression. Finally, it may increase the efficacy of cetuximab in EGFR patients with KRAS mutations.

In use, each of the drugs in the combination may be formulated as it is for monotherapy, in terms of dose size and dosage form and regimen. In this regard, the improvements resulting from their combined use may allow for the use of slightly reduced dose sizes or frequencies, as demonstrated in appropriate clinical trials.

In this method, each drug is provided in a dosage form comprising a pharmaceutically acceptable carrier and in a therapeutically effective amount. As used herein, "pharmaceutically acceptable carrier" means any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible and useful in the field of protein/antibody formulations. Examples of pharmaceutically acceptable carriers include one or more of water, saline, phosphate buffered saline, dextrose, glycerol, ethanol, and the like, as well as combinations thereof. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition. Pharmaceutically acceptable carriers may also include minor amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives or buffers, which enhance the shelf life or effectiveness of the medicament. Each of the sirpa G4 fusion protein and EGFR antibody was formulated using standard of practice in the field of therapeutic formulations. Solutions suitable for intravenous administration, such as by injection or infusion, are particularly useful.

Sterile solutions can be prepared by incorporating the active compound in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by sterile microfiltration. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle which contains a basic dispersion medium and the required other ingredients enumerated above. In the case of sterile powders for the preparation thereof, vacuum drying and freeze-drying (lyophilization) are employed which yield a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

As used herein, "effective amount" refers to an amount effective to achieve a desired therapeutic result at a desired dosage and for a particular period of time. The therapeutically effective amount of each drug in the combination may vary depending on factors such as the disease state, age, sex and weight of the individual, and the ability of the drug to elicit a desired response in the recipient, etc. A therapeutically effective amount is also one in which the therapeutically beneficial effect of the agent outweighs any toxic or detrimental effect. Of course, the EGFR antibody will be formulated in an amount appropriate for patient administration, as permitted by the regulatory authorities approving it for use in humans. Thus, in use, each drug in the combination is formulated as it would be for monotherapy, in terms of dose size and dosage form and regimen. In this regard, the synergy/benefit resulting from their combined use may allow for the use of slightly reduced dose sizes or frequencies, as demonstrated in appropriately controlled clinical trials.

The sirpa Fc fusion protein may be administered to a subject by any established route of protein delivery, in particular intravenous, intradermal, intratumoral and subcutaneous injection or infusion or by nasal administration.

Subjects targeted for treatment can be identified by first confirming the presence of disease cells of at least the phenotype of CD47+ or EGFR +, and ideally both CD47+ and EGFR +, using the assays described above.

The drugs in the combination of the invention may be administered sequentially or substantially simultaneously, i.e. simultaneously or consecutively. In embodiments, the EGFR antibody is administered prior to administration of sirpa Fc. In the alternative, the EGFR antibody may be administered after or during administration of a CD47 blocker (e.g., sirpa G4). Thus, in some embodiments, a subject undergoing therapy is a subject that has been treated with one of the combination drugs (such as an EGFR antibody), and then treated with the other of the combination drugs (such as a sirpa Fc drug). Most suitably, the drugs are administered such that their activity and effect overlap in the patient being treated, i.e. the drugs are administered in combination

The dosing regimen may be adjusted to provide the optimal desired response (e.g., therapeutic response). For example, a single bolus of each drug may be administered, or several separate doses may be administered over time, or the dose may be proportionally reduced or increased as dictated by the treatment situation. Parenteral compositions in unit dosage form are particularly advantageous for ease of administration and uniformity of dosage. As used herein, "unit dosage form" refers to physically discrete units suitable as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier.

The medicaments may be formulated in combination (e.g., as a kit) such that the combination may be introduced into the recipient in one administration (e.g., one injection or one infusion bag). Alternatively, the medicaments may be combined together as separate units in a kit provided together in a single package, and combined with written instructions teaching their use in accordance with the methods of the present invention. In another embodiment, an article of manufacture is provided that contains a sirpafc drug in combination with an EGFR antibody in amounts useful for treating the disorders described herein. The article of manufacture comprises one or both drugs of the antibody drug combination of the invention, as well as a container and label. Suitable containers include, for example, bottles, vials, syringes, and test tubes. The container may be made of various materials such as glass or plastic. The container contains a composition effective to treat the condition and may have a sterile access port (e.g., the container may be an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle). A label on or associated with the container indicates that the composition is used in combination with a sirpa Fc drug according to the present disclosure to elicit an enhanced effect on CD47+ disease cells. The article of manufacture may also include a second container comprising a pharmaceutically acceptable buffer, such as phosphate buffered saline, Ringer's solution, or dextrose solution. It may also contain other materials as desired from a commercial and user standpoint, including other buffers, diluents, filters, needles, syringes, and package inserts with instructions for use.

For administration, the dosage of the sirpa Fc drug should be in the range of about 0.0001 to 100mg/kg of host body weight, and more typically 0.01 to 10mg/kg of host body weight. For example, a SIRPa Fc dose may be 0.3mg/kg body weight, 1mg/kg body weight, 3mg/kg body weight, 5mg/kg body weight, or 10mg/kg body weight or in the range of 0.1-100 mg/kg. When the CD47 blocker is a sirpa Fc fusion protein of SEQ ID No.6, the dose can be about 1ug-5mg per injection (such as intratumoral injection).

Sirpa Fc proteins exhibit negligible binding to erythrocytes. Thus, there is no need to consider the RBC "pool" when administered with a drug combination. Furthermore, sirpa-Fc fusion proteins are dedicated antagonists of sirpa-mediated signaling, because when bound thereto, sirpa-Fc fusion proteins exhibit negligible CD47 agonism. Thus, when establishing a medically useful unit dosing regimen, there is no need to take into account any irritation caused by the drug.

Administration of cetuximab to a subject typically requires a loading dose of 400mg/m2 (2 hour intravenous infusion) and a maintenance dose of 250mg/m2 per week (1 hour intravenous infusion).

The pharmaceutical combination can be used to treat a variety of CD47+ disease cells, including CD47+ disease cells that are also EGFR + and EGFR + only disease cells. These cells include in particular CD47+ cancer cells, including liquid and solid tumors. Solid tumors can be treated with the pharmaceutical combinations of the present invention to reduce their size, number, distribution, or growth rate and to control the growth of cancer stem cells. Such solid tumors include CD47+ tumors, such as carcinomas in skin (melanoma), bladder, brain, breast, lung, colon, ovary, prostate, head and neck, colorectal tissues, liver, and other tissues, and the like. In one embodiment, the pharmaceutical combination can be used to inhibit the growth or proliferation of a hematologic cancer. As used herein, "hematological cancer" refers to cancer of the blood and includes leukemia, lymphoma, myeloma, and the like. "leukemia" refers to cancer in the blood in which too many white blood cells are produced that are ineffective against infection, thereby displacing other parts of the blood, such as platelets and red blood cells. It is understood that leukemia cases are classified as acute or chronic. Some forms of leukemia may be, for example, Acute Lymphocytic Leukemia (ALL); acute Myeloid Leukemia (AML); chronic Lymphocytic Leukemia (CLL); chronic Myelogenous Leukemia (CML); myeloproliferative disorder/neoplasm (MPDS); and myelodysplastic syndrome. "lymphoma" may refer to hodgkin's lymphoma, indolent and aggressive non-hodgkin's lymphoma, Cutaneous T Cell Lymphoma (CTCL), Peripheral T Cell Lymphoma (PTCL), burkitt's lymphoma, Mantle Cell Lymphoma (MCL), and follicular lymphoma (small and large cells), among others, including DLBCL and FL. Myelomas include Multiple Myeloma (MM), giant cell myeloma, heavy chain myeloma, and light chain and Bence-Jones myeloma.

In some embodiments, the hematologic cancer treated with the pharmaceutical combination is CD47+ leukemia, preferably selected from acute lymphocytic leukemia, acute myelocytic leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, and myelodysplastic syndrome, preferably human acute myelocytic leukemia.

In other embodiments, the hematologic cancer treated with the pharmaceutical combination is CD47+ lymphoma or bone tumor marrow selected from hodgkin's lymphoma, indolent and aggressive non-hodgkin's lymphoma, diffuse large cell lymphoma (DLBCL), mantle cell lymphoma, T cell lymphoma (including mycosis fungoides), Sezary syndrome, burkitt's lymphoma, follicular lymphoma (small and large cells), Multiple Myeloma (MM), giant cell myeloma, heavy chain myeloma and light chain or Bence-Jones myeloma, and leiomyosarcoma. When the cancer is a carcinoma, the disease may include merkel cell carcinoma, squamous cell carcinoma, and soft tissue carcinoma.

In a specific embodiment, the cancer treated with the combination of the invention is multiple myeloma. In another specific embodiment, the targeted cancer is mantle cell lymphoma. In another embodiment, the cancer treated with the combination of the invention is relapsed or refractory hodgkin lymphoma. In another specific embodiment, the CD47 blocker is sirpafc. In another specific embodiment, the EGFR antibody is cetuximab.

In some embodiments, the cancer treated is an EGFR + cancer, such as colorectal cancer, head and neck cancer, lung cancer, breast cancer, and glioma, as well as any other cancer indicated for treatment with an EGFR antibody.

In still other embodiments, cetuximab is used in combination with sirpa Fc, such as SEQ ID No.6 or SEQ ID No.7, such as for the treatment of cutaneous T cell lymphoma or multiple myeloma. In another embodiment, the combination is used to treat a T-cell lymphoma, such as mycosis fungoides or Sezary syndrome.

Thus, in a specific embodiment, there is provided the use of a CD47 blocker in combination with an EGFR antibody for the treatment of a specific CD47+ cancer, wherein:

i) the CD47 blocker is sirpa G4 of SEQ ID No.6 and the EGFR antibody is cetuximab, such as for the treatment of cancer that is head and neck cancer, colorectal cancer, cutaneous T-cell lymphoma, or multiple myeloma, or relapsed or refractory hodgkin lymphoma;

ii) the CD47 blocker is sirpa G1 of SEQ ID No.7 and the EGFR antibody is cetuximab, such as for the treatment of cancer that is head and neck cancer, colorectal cancer, cutaneous T-cell lymphoma or multiple myeloma or relapsed or refractory hodgkin lymphoma;

iii) the CD47 blocker is sirpa G4 of SEQ ID No.6 and the EGFR antibody is panitumumab, such as for the treatment of cancer as head and neck cancer, colorectal cancer, cutaneous T cell lymphoma or multiple myeloma.

It is understood that other SIRPaFc-based blockers may be used in combination with EGFR antibodies. The desired combination will exhibit a statistically significant improved cancer cell response. This may be demonstrated due to EGFR antibody activity that in combination with a CD47 blocker results in a statistically significant improvement, and vice versa, where the statistical significance is shown in the examples below, and desirably provides a p-value of >0.05, and more desirably provides a p-value of >0.01, such as > 0.001.

Combination therapy, including CD47 blockade and EGFR inhibition, may also be utilized with any other agent or modality that may be used to treat the targeted indication (such as surgery in adjuvant therapy) or with additional chemotherapy in neoadjuvant therapy.

The following non-limiting examples illustrate the present disclosure.

Referring to FIG. 1, 5X10 in Matrigel⁶Individual Fadu cells were transplanted subcutaneously on day 0 into the right side of NOD SCID (n-8 mice/group). On day 3, mice were randomly assigned and received an Intraperitoneal (IP) injection of sirpa G410 mg/kg, 5 times per week; and/or cetuximab 3mg/kg, administered 4 times every other day; or vehicle 5 times/week. (fig. 1A) shows mean tumor volume versus standard deviation for each treatment group: SIRP alpha G410 mg/kg, 5 times per week (black circle)) (ii) a And/or cetuximab (3mg/kg, administered 4 times every other day); or vehicle 5 times/week (grey squares). The curve ends when more than 25% of the animals in each group are killed. Statistical significance was calculated by one-way ANOVA (Tukey multiple comparison test) based on tumor volume at day 26. (FIG. 1B) also shows enhanced survival of tumor-bearing mice for each treatment group: SIRP α G410 mg/kg, 5 times/week (dotted line); cetuximab 3mg/kg, administered 4 times every other day (dotted line); sirpa G410 mg/kg, 5 times/week and cetuximab 3mg/kg, administered 4 times every other day (dotted line); or vehicle 5 times/week (solid line). Statistical significance of survival curves was calculated by LogRank test using Prism GraphPad software.

While the invention has been described with reference to what are presently considered to be the preferred embodiments, it is to be understood that the invention is not limited to the disclosed embodiments. To the contrary, the invention is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

All publications, patents and patent applications are herein incorporated by reference in their entirety to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference in its entirety.

Sequence listing

<110> Trillium Therapeutics Inc. (Trillium Therapeutics Inc.)

<120> improvement of CD47 blocking therapy by EGFR antibodies

<130> 9579-P55707PC00

<150> US 62/642,129

<151> 2018-03-13

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50 55 60

Glu Ser Thr Lys Arg Glu Asn Met Asp Phe Ser Ile Ser Ile Ser Asn

65 70 75 80

Ile Thr Pro Ala Asp Ala Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys

85 90 95

Gly Ser Pro Asp Thr Glu Phe Lys Ser Gly Ala Gly Thr Glu Leu Ser

100 105 110

Val Arg Ala Lys Pro Ser

115

<210> 3

<211> 229

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 3

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Ser Cys Pro Ala Pro Glu Phe

1 5 10 15

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

20 25 30

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

35 40 45

Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val

50 55 60

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser

65 70 75 80

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

85 90 95

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser

100 105 110

Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

115 120 125

Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln

130 135 140

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

145 150 155 160

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

165 170 175

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu

180 185 190

Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser

195 200 205

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

210 215 220

Leu Ser Leu Gly Lys

225

<210> 4

<211> 227

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 4

Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly

1 5 10 15

Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met

20 25 30

Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His

35 40 45

Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val

50 55 60

His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr

65 70 75 80

Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly

85 90 95

Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile

100 105 110

Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val

115 120 125

Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser

130 135 140

Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu

145 150 155 160

Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro

165 170 175

Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val

180 185 190

Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met

195 200 205

His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser

210 215 220

Pro Gly Lys

225

<210> 5

<211> 229

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 5

Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe

1 5 10 15

Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr

20 25 30

Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val

35 40 45

Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val

50 55 60

Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser

65 70 75 80

Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu

85 90 95

Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser

100 105 110

Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro

115 120 125

Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln

130 135 140

Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala

145 150 155 160

Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr

165 170 175

Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu

180 185 190

Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser

195 200 205

Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser

210 215 220

Leu Ser Leu Gly Lys

225

<210> 6

<211> 347

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 6

Glu Glu Glu Leu Gln Val Ile Gln Pro Asp Lys Ser Val Ser Val Ala

1 5 10 15

Ala Gly Glu Ser Ala Ile Leu His Cys Thr Val Thr Ser Leu Ile Pro

20 25 30

Val Gly Pro Ile Gln Trp Phe Arg Gly Ala Gly Pro Ala Arg Glu Leu

35 40 45

Ile Tyr Asn Gln Lys Glu Gly His Phe Pro Arg Val Thr Thr Val Ser

50 55 60

Glu Ser Thr Lys Arg Glu Asn Met Asp Phe Ser Ile Ser Ile Ser Asn

65 70 75 80

Ile Thr Pro Ala Asp Ala Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys

85 90 95

Gly Ser Pro Asp Thr Glu Phe Lys Ser Gly Ala Gly Thr Glu Leu Ser

100 105 110

Val Arg Ala Lys Pro Ser Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro

115 120 125

Cys Pro Ala Pro Glu Phe Leu Gly Gly Pro Ser Val Phe Leu Phe Pro

130 135 140

Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr

145 150 155 160

Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val Gln Phe Asn

165 170 175

Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg

180 185 190

Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val

195 200 205

Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser

210 215 220

Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys

225 230 235 240

Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu

245 250 255

Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe

260 265 270

Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu

275 280 285

Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe

290 295 300

Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp Gln Glu Gly

305 310 315 320

Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr

325 330 335

Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys

340 345

<210> 7

<211> 345

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 7

Glu Glu Glu Leu Gln Val Ile Gln Pro Asp Lys Ser Val Ser Val Ala

1 5 10 15

Ala Gly Glu Ser Ala Ile Leu His Cys Thr Val Thr Ser Leu Ile Pro

20 25 30

Val Gly Pro Ile Gln Trp Phe Arg Gly Ala Gly Pro Ala Arg Glu Leu

35 40 45

Ile Tyr Asn Gln Lys Glu Gly His Phe Pro Arg Val Thr Thr Val Ser

50 55 60

Glu Ser Thr Lys Arg Glu Asn Met Asp Phe Ser Ile Ser Ile Ser Asn

65 70 75 80

Ile Thr Pro Ala Asp Ala Gly Thr Tyr Tyr Cys Val Lys Phe Arg Lys

85 90 95

Gly Ser Pro Asp Thr Glu Phe Lys Ser Gly Ala Gly Thr Glu Leu Ser

100 105 110

Val Arg Ala Lys Pro Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro

115 120 125

Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys

130 135 140

Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val

145 150 155 160

Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr

165 170 175

Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu

180 185 190

Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His

195 200 205

Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys

210 215 220

Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln

225 230 235 240

Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu

245 250 255

Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro

260 265 270

Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn

275 280 285

Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu

290 295 300

Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val

305 310 315 320

Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln

325 330 335

Lys Ser Leu Ser Leu Ser Pro Gly Lys

340 345

<210> 8

<211> 449

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 8

Gln Val Gln Leu Lys Gln Ser Gly Pro Gly Leu Val Gln Pro Ser Gln

1 5 10 15

Ser Leu Ser Ile Thr Cys Thr Val Ser Gly Phe Ser Leu Thr Asn Tyr

20 25 30

Gly Val His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Leu

35 40 45

Gly Val Ile Trp Ser Gly Gly Asn Thr Asp Tyr Asn Thr Pro Phe Thr

50 55 60

Ser Arg Leu Ser Ile Asn Lys Asp Asn Ser Lys Ser Gln Val Phe Phe

65 70 75 80

Lys Met Asn Ser Leu Gln Ser Asn Asp Thr Ala Ile Tyr Tyr Cys Ala

85 90 95

Arg Ala Leu Thr Tyr Tyr Asp Tyr Glu Phe Ala Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ala Ala Ser Thr Lys Gly Pro Ser Val Phe

115 120 125

Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Pro Lys Ser Cys Asp Lys

210 215 220

Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro

225 230 235 240

Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser

245 250 255

Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp

260 265 270

Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn

275 280 285

Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val

290 295 300

Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu

305 310 315 320

Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys

325 330 335

Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr

340 345 350

Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr

355 360 365

Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu

370 375 380

Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu

385 390 395 400

Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys

405 410 415

Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu

420 425 430

Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly

435 440 445

Lys

<210> 9

<211> 213

<212> PRT

<213> Intelligent (Homo sapiens)

<400> 9

Asp Ile Leu Leu Thr Gln Ser Pro Val Ile Leu Ser Val Ser Pro Gly

1 5 10 15

Glu Arg Val Ser Phe Ser Cys Arg Ala Ser Gln Ser Ile Gly Thr Asn

20 25 30

Ile His Trp Tyr Gln Gln Arg Thr Asn Gly Ser Pro Arg Leu Leu Ile

35 40 45

Lys Tyr Ala Ser Glu Ser Ile Ser Gly Ile Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Ser Ile Asn Ser Val Glu Ser

65 70 75 80

Glu Asp Ile Ala Asp Tyr Tyr Cys Gln Gln Asn Asn Asn Trp Pro Thr

85 90 95

Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu

210

Claims

Use of a sirpa Fc protein in combination with an Epidermal Growth Factor Receptor (EGFR) antibody for treating a subject presenting with CD47+ disease cells.
2. The use of claim 1, wherein the EGFR antibody is cetuximab.
3. The use of claim 1, wherein the EGFR antibody is
4. The use of claim 1, wherein the EGFR antibody is panitumumab.
5. The use of claim 1, wherein the EGFR antibody is
6. The use of any one of claims 1-5, wherein the SIRPa Fc medicament comprises SEQ ID No. 6.
7. The use of any one of claims 1-5, wherein the SIRPa Fc medicament comprises SEQ ID No. 7.
8. The use of any one of claims 1-11, wherein the CD47+ disease cell is a cancer cell.
9. The use of claim 8, wherein the cancer cell is a hematological cancer cell or a solid tumor cell, optionally a head and neck cancer cell or a colorectal cancer cell.
10. The use of claim 9, wherein the cancer cell is a hematologic cancer cell.
11. The use of claim 10, wherein the hematological cancer cell is leukemia, lymphoma or myeloma.
12. The use according to claim 11, wherein leukemia is selected from Acute Lymphocytic Leukemia (ALL); acute Myeloid Leukemia (AML); chronic Lymphocytic Leukemia (CLL) and Chronic Myelogenous Leukemia (CML);
13. the use of claim 12, wherein the cancer is a lymphoma selected from hodgkin's lymphoma, indolent and aggressive non-hodgkin's lymphoma, burkitt's lymphoma and follicular lymphoma (small and large cells).
14. The use of claim 12, wherein the cancer is a myeloma selected from Multiple Myeloma (MM), giant cell myeloma, heavy chain myeloma, light chain or Bence-Jones myeloma, myeloproliferative disorder/neoplasm (MPDS), and myelodysplastic syndrome.
15. The use of any one of claims 1-14, wherein the EGFR antibody is administered to a subject who has received the sirpafc drug.
16. The use of any preceding claim, wherein the disease cells are CD47+ and EGFR +.
17. A pharmaceutical combination comprising an effective amount of a sirpa Fc drug and an effective amount of an EGFR antibody.
18. The combination of claim 17, wherein the EGFR antibody is cetuximab.
19. The combination of claim 16, wherein the sirpafc drug comprises SEQ ID No. 6.
20. The combination of claim 16, wherein the sirpafc drug comprises SEQ ID No. 7.
21. A kit comprising a combination according to any one of claims 16 to 20 and written instructions for the use of the combination to treat a subject presenting CD47+ disease cells.
22. A method for treating a subject having disease cells that present CD47+ and EGFR +, comprising administering to the subject a combination of sirpafc and EGFR antibodies.